本文選題：乳腺癌 + 雌激素受體α��； 參考：《浙江省醫(yī)學(xué)科學(xué)院》2017年碩士論文

【摘要】：[背景]:乳腺癌是女性常見(jiàn)腫瘤,近年來(lái)發(fā)病率顯著增高。約86%的乳腺癌是激素依賴(lài)性疾病,內(nèi)分泌治療是乳腺癌重要的治療手段。他莫昔芬是目前臨床上使用最廣泛有效的選擇性雌激素受體調(diào)節(jié)劑,但其耐藥性是臨床應(yīng)用后期的瓶頸。蛋白降解靶向嵌合體技術(shù)是將目標(biāo)蛋白和細(xì)胞內(nèi)的泛素連接酶結(jié)合,從而導(dǎo)致目標(biāo)蛋白泛素化降解。嵌合體包括三部分:一頭是靶向目標(biāo)蛋白的小分子化合物結(jié)構(gòu);另一頭是可以招募蛋白降解體系比如E3 Ligase的結(jié)構(gòu);中間通過(guò)合適的linker連接。利用蛋白降解靶向嵌合體技術(shù)為克服靶向治療藥物臨床應(yīng)用耐藥性問(wèn)題提供了新的治療策略。[目的]:篩選出高結(jié)合力、高選擇性的雌激素受體α的配體小分子化合物結(jié)構(gòu),進(jìn)行初步抗腫瘤活性研究,為進(jìn)一步合成可以同時(shí)抑制和靶向泛素化降解雌激素受體雙重藥效機(jī)制的嵌合體藥物奠定基礎(chǔ)。[方法]:以熒光偏振技術(shù)檢測(cè)候選小分子化合物與雌激素受體α及β亞型的結(jié)合力及選擇性,以雙熒光素酶報(bào)告基因檢測(cè)系統(tǒng)檢測(cè)候選小分子化合物對(duì)雌激素受體α轉(zhuǎn)錄活性的影響,以免疫組織化學(xué)法以及Western Blot鑒定乳腺癌MCF-7細(xì)胞雌激素受體α的表達(dá),以WST-1法檢測(cè)候選小分子化合物對(duì)MCF-7細(xì)胞體外增殖的影響,以乳腺癌細(xì)胞裸小鼠皮下移植瘤模型觀(guān)察候選小分子化合物對(duì)乳腺癌腫瘤體內(nèi)生長(zhǎng)的影響。[結(jié)果]:候選小分子化合物中4g和5g與雌激素受體α和β有較強(qiáng)結(jié)合力,其中化合物4g對(duì)雌激素受體α亞型的選擇性強(qiáng)于他莫昔芬;雙熒光素酶報(bào)告基因檢測(cè)系統(tǒng)檢測(cè)結(jié)果表明小分子化合物4g對(duì)雌激素受體α具有拮抗作用,對(duì)雌激素受體α表達(dá)陽(yáng)性的人乳腺癌MCF-7細(xì)胞具有顯著的增殖抑制作用,對(duì)接種MCF-7細(xì)胞的裸小鼠體內(nèi)移植瘤具有顯著的生長(zhǎng)抑制作用。[結(jié)論]:候選小分子化合物4g對(duì)雌激素受體α具有較好的結(jié)合力和選擇性,同時(shí)對(duì)乳腺癌具有抗腫瘤活性,是較為理想的構(gòu)建具有同時(shí)抑制和靶向泛素化降解雌激素受體α嵌合體的小分子化合物結(jié)構(gòu)。
[Abstract]:Background: breast cancer is a common tumor in women. The incidence of breast cancer has increased significantly in recent years. About 86% of breast cancer is hormone-dependent disease, endocrine therapy is an important treatment for breast cancer. Tamoxifen is the most widely used selective estrogen receptor modulator in clinical practice, but its resistance is the bottleneck in the later stage of clinical application. Protein degradation targeting chimera technology is to combine the target protein with the intracellular ubiquitin ligase, which leads to the degradation of the target protein ubiquitin. The chimera consists of three parts: one is the structure of small molecule which targets the target protein; the other is the structure of protein degradation system such as E3 Ligase; the middle is connected by suitable linker. The use of protein-degrading targeted chimerism provides a new therapeutic strategy for overcoming drug resistance in clinical application of targeted drugs. [objective]: to screen small ligand structures of estrogen receptor 偽 with high binding ability and selectivity, and to study its antitumor activity. It provides a basis for the further synthesis of chimeric drugs which can simultaneously inhibit and target the degradation of estrogen receptor by Ubiquitin. [methods]: the binding force and selectivity of candidate small molecular compounds with estrogen receptor 偽 and 尾 subtypes were detected by fluorescence polarization technique. Double luciferase reporter gene detection system was used to detect the effect of candidate small molecule compounds on the transcriptional activity of estrogen receptor 偽, and the expression of estrogen receptor 偽 in breast cancer MCF-7 cells was identified by immunohistochemical method and Western Blot. The effect of candidate small molecule compounds on the proliferation of MCF-7 cells in vitro was detected by WST-1 assay, and the effect of candidate small molecule compounds on the growth of breast cancer tumors in vivo was observed by subcutaneous transplantation of breast cancer cells in nude mice. [results]: among candidate small molecular compounds, 4 g and 5 g had strong binding ability with estrogen receptor 偽 and 尾, and the selectivity of compound 4 g to estrogen receptor 偽 subtype was stronger than that of tamoxifen. The results of double luciferase reporter gene detection system showed that small molecular compound 4 g had antagonistic effect on estrogen receptor 偽 and significantly inhibited the proliferation of human breast cancer MCF-7 cells with positive expression of estrogen receptor 偽. It can inhibit the growth of xenografts in nude mice inoculated with MCF-7 cells. [conclusion]: candidate small molecule compound 4 g has good binding ability and selectivity to estrogen receptor 偽, and has antitumor activity to breast cancer. It is an ideal structure of small molecular compounds with inhibition and targeting of Ubiquitin degradation of estrogen receptor 偽 chimerism.
【學(xué)位授予單位】：浙江省醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R979.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 趙晨暉;余騰驊;涂剛;;雌激素受體GPER在乳腺癌中的研究現(xiàn)狀[J];腫瘤防治研究;2015年12期

2 劉瑞磊;陳曉萍;龍梅s，

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